Preparation of hapten and monoclonal antibody of hesperetin and establishment of enzyme-linked immunosorbent assay.

Hesperetin is the aglycone of hesperidin and is widely found in the Rutaceae plants and herbal medicines. It exhibits antioxidant, detoxifying, anti-inflammatory, and antimicrobial properties, similar to hesperidin. It has also shown potential in the regulation of certain diseases. In order to detect hesperetin in complex matrix samples such as citrus and herbal, we developed an anti-hesperetin monoclonal antibody and established an indirect competitive enzyme-linked immunosorbent assay (icELISA). The half maximal inhibitory concentration (IC) was determined to be 2.03 ng/mL, the detection range was 0.39-12.73 ng/mL. In practical sample testing, the concentration of hesperidin measured by icELISA is consistent with the result of UPLC-MS/MS, and the correlation coefficient (R) is 0.97. These results showed that the established method has good accuracy, reproducibility and broad application prospects, and can be used for the detection of hesperetin in complex matrix samples (such as citrus and herbal samples).